scholarly journals A Silicone Membrane for In Vitro Feeding of Ixodes scapularis (Ixodida: Ixodidae)

2014 ◽  
Vol 51 (4) ◽  
pp. 878-879 ◽  
Author(s):  
John J. Andrade ◽  
Guang Xu ◽  
Stephen M. Rich
Keyword(s):  
Ixodes Scapularis ◽  
Silicone Membrane ◽  
In Vitro Feeding

scholarly journals Plectin ensures intestinal epithelial integrity and protects colon against colitis

2021 ◽  
Vol 14 (3) ◽  
pp. 691-702
Author(s):  
Alzbeta Krausova ◽  
Petra Buresova ◽  
Lenka Sarnova ◽  
Gizem Oyman-Eyrilmez ◽  
Jozef Skarda ◽  
...  
Keyword(s):  
Mechanical Stability ◽  
Intestinal Barrier ◽  
Intercellular Junctions ◽  
Protein Profiling ◽  
Cell Junctions ◽  
Intestinal Barrier Function ◽  
Intestinal Epithelial ◽  
Critical Determinant ◽  
In Vitro Feeding

AbstractPlectin, a highly versatile cytolinker protein, provides tissues with mechanical stability through the integration of intermediate filaments (IFs) with cell junctions. Here, we hypothesize that plectin-controlled cytoarchitecture is a critical determinant of the intestinal barrier function and homeostasis. Mice lacking plectin in an intestinal epithelial cell (IEC; PleΔIEC) spontaneously developed colitis characterized by extensive detachment of IECs from the basement membrane (BM), increased intestinal permeability, and inflammatory lesions. Moreover, plectin expression was reduced in the colons of ulcerative colitis (UC) patients and negatively correlated with the severity of colitis. Mechanistically, plectin deficiency in IECs led to aberrant keratin filament (KF) network organization and the formation of dysfunctional hemidesmosomes (HDs) and intercellular junctions. In addition, the hemidesmosomal α6β4 integrin (Itg) receptor showed attenuated association with KFs, and protein profiling revealed prominent downregulation of junctional constituents. Consistent with the effects of plectin loss in the intestinal epithelium, plectin-deficient IECs exhibited remarkably reduced mechanical stability and limited adhesion capacity in vitro. Feeding mice with a low-residue liquid diet that reduced mechanical stress and antibiotic treatment successfully mitigated epithelial damage in the PleΔIEC colon.

scholarly journals Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick—Pathogen Interactions

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 70
Author(s):  
Lourdes Mateos-Hernández ◽  
Natália Pipová ◽  
Eléonore Allain ◽  
Céline Henry ◽  
Clotilde Rouxel ◽  
...  
Keyword(s):  
Cell Line ◽  
Peripheral Nerves ◽  
Ixodes Scapularis ◽  
Embryonic Cell ◽  
Cell Subpopulation ◽  
In Vivo Experiments

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick–pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick–pathogen interactions.

scholarly journals An efficient microinjection method to generate human anaplasmosis agent Anaplasma phagocytophilum-infected ticks

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Vikas Taank ◽  
Ellango Ramasamy ◽  
Hameeda Sultana ◽  
Girish Neelakanta
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Ixodes Scapularis ◽  
In Vitro Cultures ◽  
Transmission Dynamics ◽  
Tick Feeding ◽  
Nymphal Stage ◽  
Bacterial Colony ◽  
Obligate Intracellular ◽  
Transstadial Transmission

Abstract Ticks are important vectors that transmit several pathogens including human anaplasmosis agent, Anaplasma phagocytophilum. This bacterium is an obligate intracellular rickettsial pathogen. An infected reservoir animal host is often required for maintenance of this bacterial colony and as a source for blood to perform needle inoculations in naïve animals for tick feeding studies. In this study, we report an efficient microinjection method to generate A. phagocytophilum-infected ticks in laboratory conditions. The dense-core (DC) form of A. phagocytophilum was isolated from in vitro cultures and injected into the anal pore of unfed uninfected Ixodes scapularis nymphal ticks. These ticks successfully transmitted A. phagocytophilum to the murine host. The bacterial loads were detected in murine blood, spleen, and liver tissues. In addition, larval ticks successfully acquired A. phagocytophilum from mice that were previously infected by feeding with DC-microinjected nymphal ticks. Transstadial transmission of A. phagocytophilum from larvae to nymphal stage was also evident in these ticks. Taken together, our study provides a timely, rapid, and an efficient method not only to generate A. phagocytophilum-infected ticks but also provides a tool to understand acquisition and transmission dynamics of this bacterium and perhaps other rickettsial pathogens from medically important vectors.

scholarly journals Tick populations from endemic and non-endemic areas in Germany show differential susceptibility to TBEV

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Katrin Liebig ◽  
Mathias Boelke ◽  
Domenic Grund ◽  
Sabine Schicht ◽  
Andrea Springer ◽  
...  
Keyword(s):  
Differential Susceptibility ◽  
Feeding Rates ◽  
Infection Rates ◽  
Lower Saxony ◽  
Tick Borne Encephalitis ◽  
The Central Nervous System ◽  
Tick Borne Encephalitis Virus ◽  
In Vitro Feeding ◽  
Tbev Infection

Abstract Tick-borne encephalitis virus (TBEV) is endemic in twenty-seven European countries, transmitted via the bite of an infected tick. TBEV is the causative agent of one of the most important viral diseases of the central nervous system (CNS). In Germany, 890 human cases were registered between the years 2018–2019. The castor bean tick, Ixodes ricinus, is the TBEV vector with the highest importance in Central Europe, including Germany. Despite the nationwide distribution of this tick species, risk areas of TBEV are largely located in Southern Germany. To increase our understanding of TBEV-tick interactions, we collected ticks from different areas within Germany (Haselmühl/Bavaria, Hanover/Lower Saxony) and infected them via an in vitro feeding system. A TBEV isolate was obtained from an endemic focus in Haselmühl. In two experimental series conducted in 2018 and 2019, ticks sampled in Haselmühl (TBEV focus) showed higher artificial feeding rates, as well as higher TBEV infections rates than ticks from the non-endemic area (Hanover). Other than the tick origin, year and month of the infection experiment as well as co-infection with Borrelia spp., had a significant impact on TBEV Haselmühl infection rates. Taken together, these findings suggest that a specific adaptation of the tick populations to their respective TBEV virus isolates or vice versa, leads to higher TBEV infection rates in those ticks. Furthermore, co-infection with other tick-borne pathogens such as Borrelia spp. can lower TBEV infection rates in specific populations.

scholarly journals In vitro feeding assays for hard ticks

2007 ◽  
Vol 23 (9) ◽  
pp. 445-449 ◽  
Author(s):  
Thomas Kröber ◽  
Patrick M. Guerin
Keyword(s):  
Hard Ticks ◽  
In Vitro Feeding

scholarly journals Putative morphology of Neoehrlichia mikurensis in salivary glands of Ixodes ricinus

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jaroslav Ondruš ◽  
Pavel Kulich ◽  
Oldřich Sychra ◽  
Pavel Široký
Keyword(s):  
Salivary Glands ◽  
Ixodes Ricinus ◽  
Mammalian Cells ◽  
Intracellular Pathogen ◽  
Transmission Electron ◽  
Ultrathin Sections ◽  
In Vitro Feeding ◽  
Neoehrlichia Mikurensis ◽  
Electron Lucent

Abstract Neoehrlichia mikurensis is an emerging tick-borne intracellular pathogen causing neoehrlichiosis. Its putative morphology was described in mammalian, but not in tick cells. In this study, we aim to show the presumptive morphology of N. mikurensis in salivary glands of engorged females of Ixodes ricinus. To accomplish this, we collected I. ricinus ticks in a locality with a high N. mikurensis prevalence, allowed them to feed in the artificial in vitro feeding system, dissected salivary glands and screened them by PCR for N. mikurensis and related bacteria. Ultrathin sections of salivary glands positive for N. mikurensis but negative for other pathogens were prepared and examined by transmission electron microscopy. We observed two individual organisms strongly resembling N. mikurensis in mammalian cells as described previously. Both bacteria were of ovoid shape between 0.5–0.8 μm surrounded by the inner cytoplasmic and the rippled outer membrane separated by an irregular electron-lucent periplasmic space. Detection of N. mikurensis in salivary glands of I. ricinus suggests that this bacterium uses the “salivary pathway of transmission” to infect mammals.

scholarly journals Comparison of synthetic membranes in the development of an in vitro feeding system for Dermanyssus gallinae

2009 ◽  
Vol 100 (2) ◽  
pp. 127-132 ◽  
Author(s):  
D.W.J. Harrington ◽  
J.H. Guy ◽  
K. Robinson ◽  
O.A.E. Sparagano
Keyword(s):  
Feeding Rate ◽  
Storage Conditions ◽  
Dermanyssus Gallinae ◽  
Skin Extract ◽  
Proof Of Concept ◽  
Ethical Considerations ◽  
Poultry Red Mite ◽  
Synthetic Membranes ◽  
In Vitro Feeding

AbstractAlthough artificial feeding models for the poultry red mite (Dermanyssus gallinae) most frequently use biological membranes consisting of day-old chick skin, there are ethical considerations associated with the use of skin. The few studies reported in the literature that have investigated the use of synthetic membranes to feed D. gallinae in vitro have reported limited success. The current study describes an investigation into the use of synthetic membranes made from either Nescofilm® or rayon and silicone, used either alone or in combination with different feather or skin extracts, as well as the use of capillary tubes. In all, 12 different treatments were used, and the feeding rate of D. gallinae was compared to that of day-old chick skin. Allowing mites to feed on a membrane consisting of Nescofilm with a skin extract resulted in the highest proportion of mites feeding (32.3%), which was not significantly different to the feeding rate of mites on day-old chick skin (38.8%). This study confirms that synthetic membranes can be used to feed D. gallinae artificially. Further optimization of the membrane and mite storage conditions is still necessary, but the study demonstrates a proof of concept.

Simple In-Vitro Feeding System for Northern Fowl Mites (Acari: Macronyssidae)

1992 ◽  
Vol 85 (3) ◽  
pp. 848-852 ◽  
Author(s):  
J. F. Carroll ◽  
K. W. Young ◽  
W. A. Bruce
Keyword(s):  
Feeding System ◽  
In Vitro Feeding

scholarly journals Blood Digestion by Trypsin-Like Serine Proteases in the Replete Lyme Disease Vector Tick, Ixodes scapularis

Insects ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 201
Author(s):  
Jeremiah Reyes ◽  
Cuauhtemoc Ayala-Chavez ◽  
Arvind Sharma ◽  
Michael Pham ◽  
Andrew B. Nuss ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Blood Meal ◽  
Serine Proteases ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Blood Feeding ◽  
Eastern United States ◽  
Blood Digestion ◽  
Hemoglobin Degradation

Ixodes scapularis is the major vector of Lyme disease in the Eastern United States. Each active life stage (larva, nymph, and adult) takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). This protein-rich blood meal is the only food taken by Ixodes ticks and therefore efficient blood digestion is critical for survival. Studies in partially engorged ticks have shown that the initial stages of digestion are carried out by cathepsin proteases within acidic digestive cells. In this study, we investigated the potential role of serine proteases in blood digestion in replete ticks. RNA interference was used for functional analysis and a trypsin-benzoyl-D, L-arginine 4-nitoanilide assay was used to measure active trypsin levels. Hemoglobinolytic activity was determined in vitro, with or without a serine protease inhibitor. Our data suggest that trypsin levels increase significantly after repletion. Knockdown of serine proteases negatively impacted blood feeding, survival, fecundity, levels of active trypsin in the midgut, and resulted in lower hemoglobin degradation. Incubation of midgut extract with a trypsin inhibitor resulted in 65% lower hemoglobin degradation. We provide evidence of the serine proteases as digestive enzymes in fully engorged, replete females. Understanding the digestive profile of trypsin during blood meal digestion in I. scapularis improves our understanding of the basic biology of ticks and may lead to new methods for tick control.

scholarly journals Application of image analysis in plant pathology and forest protection

2012 ◽  
Vol 52 (No. 7) ◽  
pp. 316-323
Author(s):  
P. Čermák ◽  
P. Mazal ◽  
D. Palovčíková ◽  
L. Jankovský
Keyword(s):  
Plant Pathology ◽  
Image Analysis ◽  
Forest Protection ◽  
Research Results ◽  
Computer Based ◽  
In Vitro Feeding

By means of computer-based image analysis it was possible to accelerate and automate a number of originally time-consuming activities in plant pathology and forest protection. In 1997–2005, image analysis was applied to the measuring of the size of colonies of fungi in vitro, feeding mark area, mass measuring of the size of spores and other microscopic objects, assessment of the progress of rot in a stem etc. The paper brings particular examples of the application of image analysis including methodical procedures and examples of research results obtained using Lucia G and Quick Photo Micro programs.

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